title: Autoactivation of mouse trypsinogens is regulated by chymotrypsin C via cleavage of the autolysis loop.
abstract: Chymotrypsin C (CTRC) is a proteolytic regulator of trypsinogen autoactivation in humans. CTRC cleavage of the trypsinogen activation peptide stimulates autoactivation, whereas cleavage of the calcium binding loop promotes trypsinogen degradation. Trypsinogen mutations that alter these regulatory cleavages lead to increased intrapancreatic trypsinogen activation and cause hereditary pancreatitis. The aim of this study was to characterize the regulation of autoactivation of mouse trypsinogens by mouse Ctrc. We found that the mouse pancreas expresses four trypsinogen isoforms to high levels, T7, T8, T9, and T20. Only the T7 activation peptide was cleaved by mouse Ctrc, causing negligible stimulation of autoactivation. Surprisingly, mouse Ctrc poorly cleaved the calcium binding loop in all mouse trypsinogens. In contrast, mouse Ctrc readily cleaved the Phe-150-Gly-151 peptide bond in the autolysis loop of T8 and T9 and inhibited autoactivation. Mouse chymotrypsin B also cleaved the same peptide bond but was 7-fold slower. T7 was less sensitive to chymotryptic regulation, which involved slow cleavage of the Leu-149-Ser-150 peptide bond in the autolysis loop. Modeling indicated steric proximity of the autolysis loop and the activation peptide in trypsinogen, suggesting the cleaved autolysis loop may directly interfere with activation. We conclude that autoactivation of mouse trypsinogens is under the control of mouse Ctrc with some notable differences from the human situation. Thus, cleavage of the trypsinogen activation peptide or the calcium binding loop by Ctrc is unimportant. Instead, inhibition of autoactivation via cleavage of the autolysis loop is the dominant mechanism that can mitigate intrapancreatic trypsinogen activation.
authors.affiliation: Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, Massachusetts 02118, USA.
authors.forename: Balázs Csaba
authors.initials: BC
authors.lastname: Németh
authors.forename: Thomas
authors.initials: T
authors.lastname: Wartmann
authors.forename: Walter
authors.initials: W
authors.lastname: Halangk
authors.forename: Miklós
authors.initials: M
authors.lastname: Sahin-Tóth
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002412:Cations; D002852:Chromatography, Ion Exchange; D002918:Chymotrypsin / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D016678:Genome / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010179:Pancreas / Q000201:enzymology; D010455:Peptides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D014362:Trypsinogen / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D002412:Cations; D007527:Isoenzymes; D050505:Mutant Proteins; D010455:Peptides; D014362:Trypsinogen; D002918:Chymotrypsin; C117609:chymotrypsin B; C117569:chymotrypsin C
keywords: Chymotrypsin; Enzymes; Hereditary Pancreatitis; Pancreas; Protein Processing; Proteolytic Enzymes; Serine Protease; Trypsin; Trypsinogen; Zymogen Activation